EN   EN 
 
 
 EUROPEAN  
COMMISSION   
Brussels, 28.4.2023  
COM(2023) 217 final  
ANNEXES 1 to 8  
 
ANNEXES  
to the  
proposal for a Regulation of the European Parliament and of the Council  
on detergents and surfactants, amending Regulation (EU) 2019/1020 and repealing 
Regulation (EC) No 648/2004  
{SEC(2023)  170 final}  - {SWD(2023)  113 final}  - {SWD(2023)  114 final}  -
 {SWD(2023)  115 final}    
EN 1  EN ANNEX I  
BIODEGRADABILITY REQUIREMENTS REFERRED TO IN ARTICLE 4  
ULTIMATE  BIODEGRADABILITY  CRITERIA  AND  TEST  METHODS  FOR  
SURFACTANTS  AND  SURFACTANTS  IN DETERGENTS  
1. The reference method for laboratory testing of surfactant ultimate biodegradability in 
this Regulation is based on the EN ISO standard 14593: 1999 (CO 2 headspace test).  
2. Surfactants and surfactants contained in detergents shall be ultimately biodegradable 
as determined in accordance with the criteria laid down in point 3 .  
3. Surfactants and surfactants contained in detergents shall be considered as ultimately 
biodegradable if the y meet one of the following criteria:  
(a) the level of biodegradability (mineralisation) is at least 60 % within 28 days  
measured in accord ance with  one of the following test  methods : 
(i) EN ISO Standard 14593 : 1999  — Water quality — Evaluation of 
ultimate aerobic bi odegradability of organic compounds in aqueous 
medium — Method by analysis of inorganic carbon in sealed vessels 
(CO 2 headspace test) ; 
(ii) method C.4. -C Carbon dioxide (CO 2) Evolution  Test (Modified Sturm 
Test), described in Part C , Part IV,  of the Annex to Commission   
Regulation (EC) No 440/20081; 
(iii) method C.4 -D, manometric respirometry test, described in Part C, Part V, 
of the Annex to Regulation (EC) No 440/2008;  
(iv) method C.4-E, closed bottle test, described in Part C, Part VI, of t he 
Annex to  Regulation (EC) No 440/2008 ; 
(v) method C.4-F Ministry of International Trade and Industry, Japan  
(M.I.T.I. ) described in Part C, Part V II, of the Annex to Regulation (EC) 
No 440/2008 ; 
(vi) ISO 10708 : 1997  — Water quality — Evaluation in an aqueous medium 
of the ultimate aerobic  biodegradability of organic compounds — 
Determination of biochemical oxygen demand in a two -phase closed 
bottle test.  
(b) the level of biodegradability (mineralisation) is at least 70%  within 28 days 
measured in accord ance with  one of the following test methods:  
(i) method C.4-A DOC die -away test  described in Part C, Part II, of the 
Annex to Regulation (EC) No 440/2008 ;  
(ii) method C.4-B, modified OECD screening  test described in Par t C, Part 
III, of the Annex to Regulation (EC) No 440/2008 .  
                                                 
 
1 Commission Regulation (EC) No 440/2008 of 30 May 2008 laying down test methods pursuant to 
Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, 
Evaluation, Authorisation and Restriction of Chemicals (REACH) (OJ L 142, 31.5.2008, p. 1).   
EN 2  EN Pre-adaptation shall not be used and the 10 -day window principle shall not be 
applied in any of the test methods referred to in points (a) and ( b) .  
4. The test s referred to in point  3 shall be conducted by laboratories  meeting any of the 
following conditions:  
(a) the laboratories are complying with the principles of good laboratory practice 
provided for in Directive 2004/10/EC  of the European Parliament and of the 
Council2 or international standards  recognised as being equivalent ; 
(b) the laboratories are  accredited in accordance with the standard for laboratories 
referred to in Regulation (EC) No 765/2008 .  
 
 
                                                 
 
2 Directive 2004/10/EC of the European Parliament and of the Council of 11 February 2004 on the 
harmonisation of laws, regulations and administrative provisions relating to the  application of the 
principles of good laboratory practice and the verification of their applications for tests on chemical 
substances  (OJ L 50, 20.2.2004, p. 44 ).  
EN 3  EN ANNEX II  
REQUIREMENT S FOR DETERGENTS CONTAINING MICROORGANISMS  
REFERRED TO IN ARTICLE 5  
1. Micro-organisms intentionally added to detergents  shall comply with the following  
conditions : 
(a) shall have an American Type Culture Collection (ATCC) number, belong to a 
collection of an International Deposi tory Authority (IDA) or have had their 
DNA identified in accordance with a “Strain identification protocol” (using 
16S ribosomal DNA sequencing or an equivalent method);  
(b) shall belong to both of the following:  
(i) Risk Group I as defined by Directive 2000/5 4/EC – biological agents at 
work;  
(ii) The Qualified Presumption of Safety (QPS) list issued by the European 
Food Safety Authority (EFSA) . 
This point shall not apply to micro -organisms intentionally added to detergents 
placed on the market for research and  development purposes.  
2. The following p athogenic micro -organisms  shall not be present in any of the strains 
included in the finished product when screened using the indicated test methods or 
equivalent:  
(a) E. coli, test method ISO 16649 -3:2005 ; 
(b) Streptococcus (Enterococcus) , test method ISO 21528 -1:2004 ; 
(c) Staphylococcus aureus , test method ISO 6888 -1; 
(d) Bacillus cereus , test method ISO 7932 :2004  or ISO 21871;  
(e) Salmonella , test method ISO 6579 :2002  or ISO 19250.  
3. Intentionally added micro -organisms shall not be genet ically modified 
microorganisms.  
4. Intentionally added micro -organisms shall be, with the exception of intrinsic 
resistance, susceptible to each of the major antibiotic classes , namely  
aminoglycoside, macrolide, beta -lactam, tetracycline and fluoroquinolones , in 
accordance with the  European Committee on Antimicrobial Susceptibility 
Testing  (EUCAST ) disk diffusion method or equivalent.  
5. When placed on the market, d etergents containing micro -organisms shall have a 
standard plate count equal to or greater than 1x1 05 colony -forming units (CFU s) per 
ml in accordance with ISO 4833 -1:2014 . 
6. The minimum shelf life of a detergent containing micro -organisms shall not be lower 
than 24 months and the microbial count shall not decrease by more than 10 % every 
12 months in accordance with ISO 4833 -1:2014 . 
7. Micro -organisms contained in detergents that are placed on the market in a spray 
format shall pass the acute inhalation toxicity test in accordance with the test method 
B.2., described in Part B  of the Annex to Regulation ( EC) No 440/200 8.  
8. Detergents containing micro -organisms shall not be placed on the market in a refill 
format.   
EN 4  EN 9. All claims made by the manufacturer regarding  the actions of the micro -organisms 
contained in the product shall be supported  by third -party testing.  
10. It is prohibited to claim or suggest on the label or by any other communication that 
the detergent has an antimicrobial or disinfecting effect, unless the detergent 
complies with Regulation (EU) No 528/2012.  
11. The tests referred to in point s 2, 5, 6, 7 and 9 shall be conducted by laboratories 
meeting any of the following conditions:  
(a) the laboratories are complying with the principles of good laboratory practice 
provided for in Directive 2004/10/EC  of the European Parliament and of the 
Council3 or int ernational standards recognised as being equivalent ; 
(b) the laboratories are accredited in accordance with the standard for laboratories 
referred to in Regulation (EC) No 765/2008 .  
 
 
 
                                                 
 
3 Directive 2004/10/EC of the European Parliament and of the Council of 11 February 2004 on the 
harmonisation of laws, regulations and administrative provisions relating to the application of the 
principles of good laboratory practice and the verification of their applications for tests on chemical 
substances  (OJ L 50, 20.2.2004, p. 44 ).  
EN 5  EN ANNEX III 
LIMITATIONS  ON THE CONTENT OF PHOSPHATES AND OTHER 
PHOSPHORUS COMPOUNDS  REFERRED TO IN ARTICLE 6  
Detergent  Limitations  
Consumer laundry detergents  Shall not be placed on the market if the total 
content of phosphorus is equal to or greater 
than 0,5 grams in the recommended quantity 
of the detergent to be used in the main cycle 
of the washing process for a standard 
washing machine load as defined in Part B of 
Annex V  for hard water : 
– for ‘normally soiled’ fabrics in the 
case of heavy -duty detergents,  
– for ‘lightly soiled’ fabrics in the 
case of detergents for delicate 
fabrics . 
Consumer automatic dishwasher detergents  Shall not be placed on the market i f the total 
content of phosphorus is equal to or greater 
than 0,3 grams in the standard dosage as 
defined in Part B of Annex V . 
 
   
EN 6  EN ANNEX IV 
CONFORMITY ASSESSMENT PROCEDURE REFERRED TO IN ARTICLE 7(2)  
 
Module A - Internal production protocol  
1. Description of the module  
Internal production control is the conformity assessment procedure whereby the 
manufacturer fulfils the obligations laid down in points 2, 3 and 4, and ensures and 
declares on his or her sole responsibility that the detergent or surfactant concerned 
satisfy the requirements of this Regulation that apply to them.  
2. Technical documentation  
2.1. The manufacturer shall establish the technical documentation. The documentation 
shall make it possible to assess conformity of the detergent or surfactant  with the 
relevant requirements, and shall include an adequate analysis and assessment of the 
risks.  
2.2. The technical documentation shall specify the applicable requirements and cover, as 
far as relevant for the assessment, the design, manufacture and intended use of the 
detergent or surfactant. The technical documentation shall contain, where applicable, 
at least the following elements:  
(a) a general description of the detergent or surfactant and a description of the 
intended use;  
(b) the test reports demonstrating the compliance with Annex I and, where 
applicable, with Annexes II and III;  
(c) a list of  test methods used to demonstrate compliance with the requirements of 
this Regulation ; 
(d) results of calculations made  and examinations carried out;  
(e) an ingredient data sheet which meets the following requirements:  
(i) lists all intentionally added substances and preservatives referred to in 
Part A of Annex V;  
(ii) the common chemical name or IUPAC name and, where available, the 
INCI name, the CAS number, and the European Pharmac opoeia name, is 
given for each ingredient ; 
(iii)  all substances are listed in order of decreasing abundance by weight, and  
the list is sub-divided into the following weight percentage ranges:  
(1) 10 % or more,  
(2) 1 % or over, but less than 10 %,  
(3) 0,1 % or over, but less than 1 %,  
(4) less than 0,1 %.  
For the purposes of point (e), a perfume, an essential oil, or a colouring agent shall be 
considered to be a single component.  
3. Manufacturing   
EN 7  EN The manufacturer shall take all measures necessary so that the manufacturing p rocess 
and its monitoring ensure  compliance of the detergent or surfactant  with the technical 
documentation referred to in point 2 and with the  requirements of this Regulation  
that apply to them.  
   
EN 8  EN  
ANNEX V 
LABELLING  REQUIREMENTS  
PART A – LABELLING OF CONTENTS  
The information  to be included on the labels of detergents and surfactants made available on 
the market   
1. The weight percentage ranges  ‘less than 5 % ’, ‘5 % or over but less than 15 % ’, ‘15 
% or over but less than 30 % ’, ‘30 % and more ’, shall be used to indicate the content 
of the constituents listed below where they are added in a concentration above 0,2 % 
by weight:  
(a) phosphates,  
(b) phosphonates,  
(c) anionic surfactants,  
(d) cationic surfactants,  
(e) amphoteric surfactants,  
(f) non-ionic surfactants,  
(g) oxygen -based bleaching agents,  
(h) chlorine -based bleaching agents,  
(i) EDTA and salts thereof,  
(j) NTA (nitrilotriacetic acid) and salts thereof,  
(k) phenols and halogenated phenols,  
(l) paradichlorobenzene,  
(m) aromatic hydrocarbons,  
(n) aliphatic hydrocarbons,  
(o) halogenated hydroca rbons,  
(p) soap,  
(q) zeolites,  
(r) polycarboxylates.  
2. The following classes of constituent s, if added, shall be listed irrespective of their 
concentration:  
(a) enzymes,  
(b) micro -organisms,  
(c) optical brighteners,  
(d) perfumes.  
3. Preservatives shall be listed, using where possible the system referred to in Article 33 
of Regulation (EC) No 1223/2009, irrespective of their concentration , provided that 
they meet the following conditions :   
EN 9  EN (a) contribute to the qualification of the detergent as a treated article within the 
meaning of Article 3(1) , point (l), of Regulation (EU) No 528/2012;  
(b) are labelled on a constituent of the detergent . 
The condition listed in point (b) of the first subparagraph does not have to be met 
where preservatives  do not exceed the elicitation thresholds refer red to in point 
3.4.3.3. / table 3.4.6. of Annex I to Regulation (EC) No 1272/2008 or they no longer 
have a preservation function in the  final product  even in synergies with other 
preservatives .  
4. If added at concentrations exceeding 0,01 % by weight, th e allergenic fragrances that 
are listed in entries 45, 67-92 and [X] to [X] of Annex  III to Regulation (EC) No 
1223/2009, shall be labelled  using the system referred to in Article 33 of that 
Regulation . The first sentence shall not apply to allergenic frag rances  that meet the 
labelling thresholds under Regulation (EC) No 1272/2008 .  
5. The requirements referred to in points 1 to 4 shall  not apply to professional  
detergents and surfactants , provided that  the equivalent information to that required 
in those points is provided in section 15 of the safety data sheet drawn up in 
accordance with Article 31 of Regulation (EC) No 1907/2006 . 
6. In addition to the information listed in points 1 to 5, as applicable, t he label of 
detergents containing micro -organisms shall bear the following  information:  
(a) an indication  or a precautionary statement that the product is not to be used on 
surfaces in contact with food;  
(b) an indication of the shelf life of the product ; 
(c) use instructions or special precautions, where relevant.  
 
PART B – LABELLING OF DOSAGE INFORMATION   
The information to be included on the label of consumer laundry detergents and consumer 
automatic dishwasher detergents  
1. The label of consumer laundry detergents  shall contain  the following information:  
(a) the recommended quantities and/or dosage instructions expressed in millilitres 
or grams appropriate to a standard washing machine load, for soft, medium and 
hard water hardness levels and making provision for one or two cycle wa shing 
processes,  
(b) for heavy -duty detergents, the number of standard washing machine loads of 
‘normally soiled’ fabrics, and, for detergents for delicate fabrics, the number of 
standard washing machine loads of ‘lightly soiled’ fabrics, that can be washed 
with the contents of the package using water of medium hardness, 
corresponding to 2,5 millimoles CaCO 3/l, 
(c) the capacity of any measuring cup, if provided, shall be indicated in millilitres 
or grams, and markings shall be provided to indicate the dose of detergent 
appropriate for a standard washing machine load for soft, medium and hard 
water hardness levels,  
2. For the purposes of point 1, t he standard was hing machine loads shall be  4,5 kg dry 
fabric for heavy -duty detergents and 2 ,5 kg dry fabric for light -duty detergents. A 
detergent shall be considered to be a heavy -duty detergent unless the claims of the  
EN 10  EN manufacturer predominantly promote fabric care, namely  low temperature wash, 
delicate fibres and colours.  
3. The label of consumer automatic dishwasher det ergents shall indicate the standard 
dosage expressed in grams or millilitres  or number of tablets for the main washing 
cycle for normally soiled tableware in a fully loaded 12 place settings dishwasher, 
adjusting the standard dosage , where relevant, for soft, medium, and hard water 
hardness.  
 
PART C – DIGITAL LABELLING   
The following content information referred to in part A, may be pr ovided on the digital label 
only, in accordance with Article 16(1),  second subparagraph , in the manner specified in  that 
part:  
(a) anionic surfactants;  
(b) cationic surfactants;  
(c) amphoteric surfactants;  
(d) non-ionic surfactants;  
(e) phosphates ; 
(f) phosphonates;  
(g) soap.  
 
PART D – SIMPLIFIED DOSAGE INFORMATION FOR CONSUMER LAUNDRY 
DETERGENTS  
The simplified dosage grid shall contain the following information :  
(a) basic instructions for use, where relevant;  
(b) the recommended quantities based on medium/average water hardness and 
different degrees of fabric soiling; and  
(c) an indication o f the washing machine load.  
   
EN 11  EN ANNEX VI 
PRODUCT PASSPORT  
The product passport shall include the following information:  
(a) the unique product identifier of the detergent or surfactant;  
(b) the name, the address of the manufacturer or the manufacturer’s authorised 
representative as well the manufacturer’s unique operato r identifier;  
(c) the identification of detergent or surfactant allowing traceability, including a 
colour image of sufficient clarity to enable the ide ntification of the detergent or  
surfactant;  
(d) the commodity code under which the detergent or surfactant  is classified at the 
moment the product passport is created, as set out in Council Regulation (EEC) 
No 2658/874; 
(e) references to Union legal acts that the detergent or surfactant complies with;  
(f) a full list of substances intentionally added in the detergent or sur factant and of 
preservatives labelled in accordance with part A, point 3, first subparagraph, 
point (b), of Annex V, using the International Nomenclature of Cosmetic 
Ingredients, or where it is not available, the European Pharmacopoeia name 
and, when also the latter is not available, the common chemical name or 
International Union of Pure and Applied Chemists name . 
The obligation referred to in point (f) shall not apply to professional detergents, or to 
surfactants for professional detergents, for which a safety data sheet referred to in 
Article 31 of Regulation (EC) No 1907/2006 is available.  
 
 
                                                 
 
4 Counci l Regulation (EEC) No 2658/87 of 23 July 1987 on the tariff and statistical nomenclature and on 
the Common Customs Tariff  (OJ L 256, 7.9.1987, p. 1 ).  
EN 12  EN ANNEX VII  
TEST METHODS REFERRED TO IN ARTICLE 22(2)  
1. REFERENCE METHOD (CONFIRMATORY TEST ) 
1.1. Definition  
This method describes a laborator y model of the activated sludge and secondary 
settler which is designed to simulate municipal sewage treatment. Improved state -of-
the-art operating conditions can be applied to this test method as described in EN 
ISO 11733.  
1.2. Equipment needed for measurement  
The method of measurement employs the small -activated sludge plant shown in 
Figure 1, and in greater detail in Figure 2. The equipment consists of a sewage vessel 
A for synthetic sewage, dosing pump B, aeration vessel C, settling vessel D, air -lift 
pump E to recycle the activated sludge, and vessel F for collecting the treated 
effluent.  
Vessels A and F must be of glass or suitable plastic and hold at least twenty -four 
litres. Pump B must provide a constant flow of synthetic sewage to the aeratio n 
vessel; this vessel, during normal operation, contains three litres of mixed liquor. A 
sintered aeration cube G is suspended in the vessel C at the apex of the cone. The 
quantity of air blown through the aerator shall be monitored by means of a flow 
mete r H. 
1.3. Synthetic sewage  
A synthetic sewage is employed for the test. Dissolve in each litre of tap water:  
– 160 mg peptone;  
– 110 mg meat extract;  
– 30 mg urea, CO(NH 2)2; 
– 7 mg sodium chloride, NaCl;  
– 4 mg calcium chloride, CaCl 2.2H 2O; 
– 2 mg magnesium sulphate, MgSO 4.7H 2O; 
– 28 mg of di -potassium hydrogen phosphate, K 2HPO 4; 
– and 10 ± 1 mg of the surfactant.  
The synthetic sewage is freshly prepared daily.  
1.4. Preparation of samples  
Uncompounded surfactants are examined in the original state. Active content of 
surfac tant samples must be determined in order to prepare the synthetic sewage 
(point 1.3). 
1.5. Operation of equipment  
Initially, fill aeration vessel C and settling vessel D with synthetic sewage. The 
height of the vessel D should be so fixed that the volume c ontained in the aeration 
vessel C is three litres. Inoculation is made by introducing 3 ml of a secondary 
effluent of good quality, freshly collected from a treatment plant dealing with a  
EN 13  EN predominantly domestic  sewage. The effluent must be kept under aero bic conditions 
in the period between sampling and application. Then set the aerator G, air -lift E and 
dosing device B in operation. The synthetic sewage must pass through the aeration 
vessel C at a rate of one litre per hour; this gives a mean retention ti me of three 
hours.  
The rate of aeration should be so regulated that the contents of vessel C are kept 
constantly in suspension and the dissolved oxygen content is at least 2 mg/l. Foaming 
must be prevented by appropriate means. Anti -foaming agents that inh ibit the 
activated sludge or contain surfactants must not be used. The air -lift pump E must be 
set so that the activated sludge from the settling vessel is continually and regularly 
recycled to aeration vessel C. Sludge which has accumulated around the top  of the 
aeration vessel C, in the base of the settling vessel D, or in the circulation circuit 
must be returned to the circulation at least once each day by brushing or some other 
appropriate means. When the sludge fails to settle, its settleability may be  increased 
by the addition of 2 ml portions of a 5 % solution of ferric chloride, repeated as 
necessary.  
The effluent from the settling vessel D is accumulated in vessel F for twenty -four 
hours, following which a sample is taken after thorough mixing. Vess el F must then 
be carefully cleaned.  
1.6. Checking measuring equipment  
The surfactant content (in mg/l) of the synthetic sewage is determined immediately 
before use.  
The surfactant content (in mg/l) of the effluent collected over twenty -four hours in 
vesse l F should be determined analytically by the same method, immediately after 
collection: otherwise the samples must be preserved, preferably by freezing. The 
concentrations must be determined to the nearest 0,1 mg/l surfactant  
As a check on the efficiency o f the process, the chemical oxygen demand (COD) or 
the dissolved organic carbon (DOC) of the glass fibre filtered effluent accumulated 
in vessel F and of the filtered synthetic sewage in vessel A is measured at least twice 
per week.  
The reduction in COD or  DOC should level off when a roughly regular daily 
surfactant degradation is obtained at the end of the running -in period shown in Figure 
3. 
The content of dry matter in the activated sludge contained in the aeration vessel 
should be determined twice a wee k in g/l. If it is more than 2,5 g/l, the excess 
activated sludge must be discarded.  
The degradation test is performed at room temperature; this should be steady and 
kept between 19 -24 ° C.  
1.7. Calculation of biodegradability  
The percentage degradation of  surfactant must be calculated every day on the basis 
of the surfactant content in mg/l of the synthetic sewage and of the corresponding 
effluent accumulated in vessel F.  
The degradability values thus obtained should be presented graphically as in Figure 
3.  
EN 14  EN The degradability of the surfactant should be calculated as the arithmetic mean of the 
values obtained over the twenty -one days that follow the running -in and 
acclimatisation period, during which degradation has been regular and the operation 
of the plan t trouble -free. In any event the duration of the running -in period should 
not exceed six weeks.  
The daily degradation values are calculated to the nearest 0,1 % but the final result is 
given to the nearest whole number.  
In some cases it may be permissible to reduce the frequency of sampling but at least 
fourteen results collected over the twenty -one days which follow the running -in 
period should be used in calculating the average.  
2. DETERMINATION OF ANIO NIC SURFACTANTS IN B IODEGRADABILITY TEST S 
2.1. Princi ple 
The method is based on the fact that the cationic dye methylene blue forms blue salts 
with anionic surfactants (MBAS), which can be extracted with chloroform. To 
eliminate interference, the extraction is first effected from alkaline solution and the 
extract is then shaken with acidic methylene blue solution. The absorbency of the 
separated organic phase is measured photometrically at the wavelength of maximum 
absorption of 650 nm.  
2.2. Reagents and equipment  
2.2.1.  Buffer solution pH 10  
Dissolve 24 g so dium bicarbonate, NaHCO 3 AR, and 27 g anhydrous sodium 
carbonate (Na 2CO 3) AR in deionised water and dilute to 1000 ml.  
2.2.2.  Neutral methylene blue solution  
Dissolve 0,35 g methylene blue AR in deionised water and dilute to 1000 ml. Prepare 
the solution at least twenty -four hours before use. The absorbency of the blank 
chloroform phase, measured against chloroform must not exceed 0,015 per 1 cm of 
layer thickness at 650 nm.  
2.2.3.  Acidic methylene blue solution  
Dissolve 0,35 g methylene blue AR in 500 ml deionised water and mix with 6,5 ml 
H2SO 4 (d = 1,84 g/ml). Dilute to 1000 ml with deionised water. Prepare the solution 
at least twenty -four hours before use. The absorbency of the blank chloroform phase, 
measured against chloroform must not exceed 0,015 p er 1 cm of layer thickness at 
650 nm.  
2.2.4.  Chloroform (trichloromethane) AR freshly distilled  
2.2.5.  Dodecyl benzene sulphonic acid methyl ester  
2.2.6.  Ethanolic potassium hydroxide solution, KOH 0,1 M  
2.2.7.  Ethanol pure, C 2H5OH 
2.2.8.  sulphuric acid, H 2SO4 0,5 M  
2.2.9.  Phenolphthalein solution  
Dissolve 1 g phenolphthalein in 50 ml ethanol and add 50 ml deionised water while 
stirring continuously. Filter off any precipitate obtained.   
EN 15  EN 2.2.10.  Methanolic hydrochloric acid: 250 ml hydrochloric acid AR and 7 50 ml methanol  
2.2.11.  Separating funnel, 250 ml  
2.2.12.  Graduated flask, 50 ml  
2.2.13.  Graduated flask, 500 ml  
2.2.14.  Graduated flask, 1000 ml  
2.2.15.  Round -bottomed flask with ground glass stopper and reflux condenser, 250 ml; 
boiling granules  
2.2.16.  pH meter  
2.2.17.  Photometer for measurements at 650 nm, with 1 to 5 cm cells  
2.2.18.  Qualitative grade filter paper  
2.3. Procedure  
The samples for analysis must not be taken through a layer of foam.  
After thorough cleaning with water, the equipment used fo r the analysis must be 
thoroughly rinsed with methanolic hydrochloric acid ( point 2.2.10) and then with 
deionised water before using.  
Filter the activated sludge plant influent and effluent to be examined immediately on 
sampling. Discard the first 100 ml o f the filtrates.  
Place a measured volume of the sample, neutralised if necessary, into a 250 ml 
separating funnel ( point 2.2.11). The volume of sample should contain between 20 
and 150 g of MBAS. At the lower MBAS content, up to 100 ml of sample may be 
used. When using less than 100 ml, dilute to 100 ml with deionised water. Add to the 
sample 10 ml of buffer solution ( point 2.2.1), 5 ml of neutral methylene blue solution 
(point 2.2.2) and 15 ml of chloroform ( point 2.2.4). Shake the mixture uniformly and 
not too vigorously for one minute. After phase separation, run the chloroform layer 
into a second separating funnel, containing 110 ml of deionised water and 5 ml of 
acidic methylene blue solution ( point 2.2.3). Shake the mixture for one minute. Pass 
the c hloroform layer through a cotton -wool filter previously cleaned and wetted with 
chloroform into a graduated flask ( point 2.2.12).  
Extract the alkaline and acid solutions three times, using 10 ml of chloroform for the 
second and third extractions. Filter th e combined chloroform extracts through the 
same cotton wool filter and dilute to the mark in the 50 ml flask ( point 2.2.12) with 
chloroform used for rewashing the cotton wool. Measure the absorbency of the 
chloroform solution with a photometer at 650 nm in  1 to 5 cm cells against 
chloroform. Run a blank determination through the whole procedure.  
2.4. Calibration curve  
Prepare a calibration solution from the standard substance dodecylbenzene sulphonic 
acid methyl ester (tetrapropylene type mol. wt. 340) afte r saponification into the 
potassium salt. The MBAS is calculated as sodium dodecyl benzene sulphonate (mol. 
wt. 348).  
From a weighing pipette, weigh 400 to 450 mg of dodecyl -benzene -sulphonic -acid-
methyl -ester ( point 2.2.5) to the nearest 0,1 mg in a round -bottomed flask and add 50 
ml of ethanolic potassium hydroxide solution ( point 2.2.6) and some boiling 
granules. After mounting the reflux condenser, boil for one hour. After cooling, wash  
EN 16  EN the condenser and ground glass joint with about 30 ml of ethanol, a nd add these 
washings to the contents of the flask. Titrate the solution with sulphuric acid against 
phenolphthalein until it becomes colourless. Transfer this solution to a 1000 ml 
graduated flask ( point  2.2.14), dilute to the mark with deionised water an d mix.  
Part of this surfactant stock solution is then further diluted. Withdraw 25 ml, transfer 
to a 500 ml graduated flask ( point  2.2.13), dilute to the mark with deionised water 
and mix.  
This standard solution contains:  
 
where E is the sample weight in mg. 
To establish the calibration curve, withdraw 1, 2, 4, 6, 8 ml portions of the standard 
solution and dilute each to 100 ml with deionised water. Then proceed as stated 
under point  2.3 including a blank determination.  
2.5. Calculation of results  
The amou nt of anionic surfactant (MBAS) in the sample is read from the calibration 
curve ( point  2.4). The MBAS content of the sample is given by:  
 
where: V = ml volume of the sample used.  
Express the results as sodium dodecylbenzene sulphonate (MW 348).  
2.6. Expression of results  
Express the results as MBAS mg/l to the nearest 0,1.  
3. DETERMINATION OF NON -IONIC SURFACTANTS IN  BIODEGRADATION TEST  LIQUORS  
3.1. Principle  
Surface active agents are concentrated and isolated by gas stripping. In the sample 
used, the  quantity of non -ionic surfactant should be in the range 250 -800 g.  
The stripped surfactant is dissolved in ethyl acetate.  
After phase separation and evaporation of the solvent, the non -ionic surfactant is 
precipitated in aqueous solution with modified Dra gendorff reagent (KBiI 4 + BaCl 2 + 
glacial acetic acid).  
The precipitate is filtered, washed with glacial acetic acid and dissolved in 
ammonium tartrate solution. The bismuth in the solution is titrated potentiometrically 
with pyrrolidinedithiocarbamate sol ution at pH 4 -5 using a bright platinum indicator 
electrode and a calomel or silver/silver chloride reference electrode. The method is 
applicable to non -ionic surfactants containing 6 -30 alkylene oxide groups.  
 
EN 17  EN The titration result is multiplied by the empi rical factor of 54 for conversion to the 
reference substance nonylphenol condensed with 10 mols ethylene oxide (NP 10).  
3.2. Reagents and Equipment  
Reagents are to be made up in deionised water.  
3.2.1.  Pure ethyl acetate, freshly distilled.  
3.2.2.  Sodium bicarbonate, NaHCO 3 AR. 
3.2.3.  Dilute hydrochloric acid [20 ml concentrated acid (HCl) diluted to 1000 ml 
with water]  
3.2.4.  Methanol AR, freshly distilled, stored in a glass bottle.  
3.2.5.  Bromocresol purple, 0,1 g in 100 ml methanol.  
3.2.6.  Precipitating  agent: the precipitating agent is a mixture of two volumes of 
solution A and one volume of solution B. The mixture is stored in a brown bottle and 
can be used for up to one week after mixing.  
3.2.6.1.  Solution A  
Dissolve 1,7 g bismuth nitrate, BiONO 3.H2O AR, in 20 ml glacial acetic acid, and 
make up to 100 ml with water. Then dissolve 65 g potassium iodide AR in 200 ml 
water. Mix these two solutions in a 1000 ml measuring flask, add 200 ml glacial 
acetic acid ( point  3.2.7) and make up to 1000 ml with water . 
3.2.6.2.  Solution B  
Dissolve 290 g barium chloride, BaCl 2.2H 2O AR, in 1000 ml of water.  
3.2.7.  Glacial acetic acid 99 -100 % (lower concentrations are unsuitable).  
3.2.8.  Ammonium tartrate solution: mix 12,4 g tartaric acid AR and 12,4 ml of 
ammonia solut ion AR (d = 0,910 g/ml) and make up to 1000 ml with water (or use 
the equivalent amount of ammonium tartrate AR).  
3.2.9.  Dilute ammonia solution: 40 ml ammonia solution AR (d = 0,910 g/ml) diluted 
to 1000 ml with water.  
3.2.10.  Standard acetate buffer: dis solve 40 g solid sodium hydroxide AR, in 
500 ml water in a beaker and allow to cool. Add 120 ml glacial acetic acid ( point  
3.2.7). Mix thoroughly, cool and transfer to a 1000 ml volumetric flask. Make up to 
the mark with water.  
3.2.11.  Pyrrolidinedithiocar bamate solution (known as ‘carbate solution’): 
dissolve 103 mg sodium pyrrolidinedithiocarbamate, C 5H8NNaS 2.2H 2O, in about 500 
ml water, add 10 ml of n -amyl alcohol AR and 0,5 g NaHCO 3 AR, and make up to 
1000 ml with water.  
3.2.12.  Copper sulphate solution (for standardisation of point  3.2.11).  
STOCK SOLUTION  
Mix 1,249 g copper sulphate, CuSO 4.5H 2O AR, with 50 ml 0,5 M sulphuric 
acid and make up to 1000 ml with water.  
STANDARD  SOLUTION  
Mix 50 ml stock solution with 10 ml 0,5 M H 2SO 4 and make up to 1000 ml 
with water.   
EN 18  EN 3.2.13.  Sodium chloride AR.  
3.2.14.  Gas-stripping apparatus (see Figure 5).  The diameter of the sintered 
disc must be the same as the internal diameter of the cylinder.  
3.2.15.  Separating funnel, 250 ml.  
3.2.16.  Magnetic stirrer with magnet 25 -30 mm.  
3.2.17.  Gooch crucible, diameter of the perforated base = 25 mm, Type G4.  
3.2.18.  Circular glass -fibre filter papers, 27 mm diameter with fibre diameter 
0,3-1,5 m.  
3.2.19.  Two filter flasks with adapters and rubber collars, 50 0 and 250 ml 
respectively.  
3.2.20.  Recording potentiometer fitted with a bright platinum indicator 
electrode and a calomel or silver/silver chloride reference electrode with a 250 mV 
range, with automatic burette of 20 -25 ml capacity, or alternative manual  equipment.  
3.3. Method  
3.3.1.  Concentration and separation of the surfactant  
Filter the aqueous sample through a qualitative filter paper. Discard the first 100 ml 
of the filtrate.  
Into the stripping apparatus, previously rinsed with ethyl acetate, place a measured 
quantity of the sample, such that it contains between 250 -800 g non -ionic surfactant.  
To improve the separation add 100 g sodium chloride and 5 g sodium bicarbonate.  
If the volume of the sample exceeds 500 ml, add these salts to the stripping ap paratus 
in solid form, and dissolve by passing nitrogen or air through.  
If a smaller -sized sample is used, dissolve the salts in 400 ml water and then add to 
the stripping apparatus.  
Add water to bring the level to the upper stopcock.  
Cautiously add 100 ml  ethyl acetate on top of the water.  
Fill the wash -bottle in the gas -line (nitrogen or air) two -thirds full with ethyl acetate.  
Pass a gas stream of 30 -60 l/h through the apparatus; the use of a flowmeter is 
recommended. The rate of aeration must be increas ed gradually at the beginning. The 
gas rate must be so adjusted that the phases remain noticeably separate to minimise 
the mixing of the phases and the solution of the ethyl acetate in the water. Stop the 
gas flow after five minutes.  
If there is a reductio n of more than 20 % in the volume of the organic phase through 
solution in water, the sublation must be repeated paying special attention to the rate 
of gas flow.  
Run off the organic phase into a separating funnel. Return any water in the separating 
funnel  from the aqueous phase — it should only be a few ml — to the stripping 
apparatus. Filter the ethyl acetate phase through a dry qualitative filter paper into a 
250 ml beaker.  
Put a further 100 ml ethyl acetate into the stripping apparatus and again pass nitrogen 
or air through for five minutes. Draw off the organic phase into the separating funnel  
EN 19  EN used for the first separation, reject the aqueous phase and run the organic phase 
through the same filter as the first ethyl acetate portion. Rinse both the sep arating 
funnel and the filter with about 20 ml ethyl acetate.  
Evaporate the ethyl acetate extract to dryness using a water -bath (fume cupboard). 
Direct a gentle stream of air over the surface of the solution to accelerate the 
evaporation.  
3.3.2.  Precipitat ion and filtration  
Dissolve the dry residue from 3.3.1 in 5 ml methanol, add 40 ml water and 0,5 ml 
dilute HCl ( point  3.2.3) and stir the mixture with a magnetic stirrer.  
To this solution add 30 ml of precipitating agent ( point  3.2.6) from a measuring 
cylinder. The precipitate forms after repeated stirring. After stirring for ten minutes 
leave the mixture to stand for at least five minutes.  
Filter the mixture through a Gooch crucible, the base of which is covered with a 
glass -fibre filter paper. First wash the filter under suction with about 2 ml glacial 
acetic acid. Then thoroughly wash the beaker, magnet, and crucible with glacial 
acetic acid, of which about 40 -50 ml is necessary. It is not necessary to quantitatively 
transfer the precipitate adhering to t he sides of the beaker, to the filter, because the 
solution of the precipitate for the titration is returned to the precipitating beaker, and 
the remaining precipitate will then be dissolved.  
3.3.3.  Dissolution of the precipitate  
Dissolve the precipitate i n the filter crucible by the addition of hot ammonium 
tartrate solution (about 80 ° C) ( point  3.2.8) in three portions of 10 ml each. Allow 
each portion to stand in the crucible for some minutes before being sucked through 
the filter into the flask.  
Put th e contents of the filter flask into the beaker used for the precipitation. Rinse the 
sides of the beaker with a further 20 ml of tartrate solution to dissolve the rest of the 
precipitate.  
Carefully wash the crucible, adapter and filter flask with 150 -200 m l water, and 
return the rinsing water to the beaker used for the precipitation.  
3.3.4.  The titration  
Stir the solution using a magnetic stirrer ( point  3.2.16), add a few drops of 
bromocresol purple ( point  3.2.5) and add the dilute ammonia solution ( point  3.2.9) 
until the colour turns violet (the solution is initially weakly acid from the residue of 
acetic acid used for rinsing).  
Then add 10 ml standard acetate buffer ( point  3.2.10), immerse the electrodes in the 
solution, and titrate potentiometrically with  standard ‘carbate solution’ ( point  3.2.11), 
the burette tip being immersed in the solution.  
The titration rate should not exceed 2 ml/min.  
The endpoint is the intersection of the tangents to the two branches of the potential 
curve.  
It will be observed occ asionally that the inflection in the potential curve becomes 
flattened; this can be eliminated by carefully cleaning the platinum electrode (by 
polishing with emery paper).   
EN 20  EN 3.3.5.  Blank determinations  
At the same time run a blank determination through the whole procedure with 5 ml 
methanol and 40 ml water, according to the instructions in point 3.3.2. The blank 
titration should be below 1 ml, otherwise the purity of the reagents ( points 3.2.3, 
3.2.7, 3.2.8, 3.2.9, 3.2.10) is suspect, especially their content of heavy metals, and 
they must be replaced. The blank must be taken into account in the calculation of the 
results.  
3.3.6.  Control of the factor of the ‘carbate solution’  
Determine the factor for the carbate solution on the day of use. To do this, titrate 10 
ml of the copper sulphate solution ( point  3.2.12) with ‘carbate solution’ after the 
addition of 100 ml water and 10 ml standard acetate buffer ( point  3.2.10). If the 
amount used is a ml, the factor f is:  
 
 
and all the results of the titration a re multiplied by this factor.  
3.4. Calculation of results  
Every non -ionic surfactant has its own factor, depending on its composition, 
particularly on the length of the alkene oxide chain. The concentration of non -ionic 
surfactant is expressed in relation to a standard substance — a nonyl phenol with ten 
ethylene oxide units (NP 10) — for which the conversion factor is 0,054.  
Using this factor the amount of surfactant present in the sample is found expressed as 
mg of NP 10 equivalent, as follows:  
(b — c) xfx 0,054 = mg non -ionic surfactant as NP 10  
where:  
b = volume of ‘carbate solution’ used by the sample 
(ml),  
c = volume of ‘carbate solution’ used by the blank 
(ml),  
f = factor of the ‘carbate solution’.  
 
3.5. Expression of results  
Express the results in  mg/l as NP 10 to the nearest 0,1.  
 
 
EN 21  EN Figure 1 Activated sludge plant: overviews  
 
 
A Storage vessel  
B Dosing device  
C Aeration chamber (three litres capacity)  
D Settling vessel  
E Air-lift pump  
F Collector  
G Sintered aerator  
H Air-flow meter  
I Air 
 
 
EN 22  EN Figure 2 Activated sludge plant: detail (dimensions in millimetres)  
 
 
 
A Liquid level  
B Hard PVC  
C Glass or waterproof plastic (hard PVC)  
 
 
EN 23  EN Figure 3 Calculation of biodegradability - Confirmatory test  
 
A Running -in period  
B Period used for calculation (twenty -one days)  
C Readily biodegradable surfactant  
D Surfactant not readily biodegradable  
E Biodegradation ( %)  
F Time (days)  
 
 
EN 24  EN ANNEX VIII 
CORRELATION TABLE  
Regulation (EC) No 648/2004  This Regulation  
Article 1(1)  Article 1(1)  
Article 1(2)  - 
Article 2(1)  Article 2, point (1)  
Article 2(1a)  Article 2, point (2)  
Article 2(1b)  Article 2, point (3)  
Article 2(2) - 
Article 2(3)  Article 2, point (6 ) 
Article 2(4)  Article 2, point (7 ) 
Article 2(5)  Article 2, point (8 ) 
Article 2(6)  Article 2, point (11 ) 
Article 2(7)  - 
Article 2(8)  Article 2, point (12 ) 
Article 2(9)  Article 2, point (14 ) 
Article 2(9a)  Article 2, point (13 ) 
Article 2(10)  Article 2, point (15 ) 
Article 2(11)  - 
Article 2(12)  Article 2, point (5 ) 
Article 3(1)  Article 3(1) and Article 4(2)  
Article 3(2)  - 
Article 3(3)  Article 7(1)  
Article 4(1)  Article 4(1)  
Article 4(2)  - 
Article 4(3)  - 
Article 4a  Article 6  
Article 5(1)  - 
Article 5(2)  - 
Article 5(3)  - 
Article 5(4)  - 
Article 5(5)  - 
Article 5(6)  - 
Article 6(1)  -   
EN 25  EN Article 6(2)  - 
Article 6(3)  - 
Article 6(4)  - 
Article 7  - 
Article 8(1)  - 
Article 8(2)  - 
Article 8(3)  - 
Article 8(4) - 
Article 9(1)  Article 8(2)  
Article 9(2)  - 
Article 9(3)  Article 7( 6) 
Article 10(1)  - 
Article 10(2)  Article 22(2)  
Article 11(1)  Article 1(2), point (b)  
Article 11(2) and (3)  Article 15(3)  
Article 11(4)  Article 15(4)  
Article 11(5)  Article 15(5)  
Article 11(6)  - 
Article 12  Article 28  
Article 13  Article 26  
Article 13a(1)  Article 27(1)  
Article 13a(2)  Article 27(2)  
Article 13 a(3) Article 27(3)  
Article 13a(4)  Article 27(5)  
Article 13a(5)  Article 27(6)   
EN 26  EN Article 14(1)  Article 3(2) 
Article 14(2)  - 
Article 14(3)  - 
Article 14(4)  - 
Article 14(5)  - 
Article 15(1) , first subparagraph  Article 24(1)  
Article 15(1) , second subparagraph  Article 24(3)  
Article 15(2)  Article 25(4)  
Article 16(1)  - 
Article 16(2)  - 
Article 17  Article 33  
Article 18  Article 29  
Article 19  Article 35